Isolation of amplified and overexpressed DNA sequences from Adriamycin-resistant human breast cancer cells

C. R. Fairchild, S. P. Ivy, C. S. Kao-Shan, J. Whang-Peng, N. Rosen, M. A. Israel, P. W. Melera, K. H. Cowan, M. E. Goldsmith

研究成果: 雜誌貢獻文章

280 引文 (Scopus)

摘要

An MCF-7 human breast cancer cell line was selected which was 200-fold more resistant to Adriamycin than the wild type cell line. This Adriamycin-resistant (Adr(R)) cell line exhibited a multidrug-resistant phenotype and was cross-resistant to a wide range of antineoplastic agents including Vinca alkaloids, anthracyclines, and epipodophyllotoxins. Cytogenetic analysis of the Adr(R) cell line showed the presence of homogeneously staining regions on several chromosomes which were not present in the parental cell line. Using the technique of in-gel renaturation, DNA sequences which were amplified 50- to 100-fold in the Adr(R) cell line and which covered a total of over 140 kilobases were isolated. In addition, Adr(R) cells were found to contain amplified and overexpressed sequences which were homologous to hamster P-glycoprotein gene sequences. A hamster cDNA P-glycoprotein gene probe was used to screen a λgt10 cDNA library made from human Adr(R) cell line mRNA and human cDNA sequences homologous to the P-glycoprotein gene were isolated. Hybridization studies with the cloned human cDNA (pADR1) showed that the Adr(R) MCF-7 cell line contained a 60-fold amplification of this DNA sequence and that polyadenylated mRNA from the Adr(R) cell line contained a 4.8-kilobase transcript which was overexpressed 45-fold. There was a direct correlation between DNA and RNA copy number of this sequence and level of resistance among several MCF-7 Adriamycin-resistant cell lines. In situ hybridization studies demonstrated that the human P-glycoprotein gene sequence was found on chromosome 7q21.1 in normal human lymphocytes and that amplified DNA sequences isolated from the Adr(R) MCF-7 cells by the in-gel hybridization technique were linked to the human P-glycoprotein sequences in the homogeneously staining regions in the Adr(R) cells.
原文英語
頁(從 - 到)5141-5148
頁數8
期刊Cancer Research
47
發行號19
出版狀態已發佈 - 一月 1 1987
對外發佈Yes

指紋

Doxorubicin
Breast Neoplasms
Cell Line
P-Glycoprotein
Complementary DNA
MCF-7 Cells
Cricetinae
Genes
Chromosomes
Gels
Podophyllotoxin
Vinca Alkaloids
Staining and Labeling
Messenger RNA
Cytogenetic Analysis
Anthracyclines
Sequence Homology
Gene Library
Antineoplastic Agents
In Situ Hybridization

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

引用此文

Fairchild, C. R., Ivy, S. P., Kao-Shan, C. S., Whang-Peng, J., Rosen, N., Israel, M. A., ... Goldsmith, M. E. (1987). Isolation of amplified and overexpressed DNA sequences from Adriamycin-resistant human breast cancer cells. Cancer Research, 47(19), 5141-5148.

Isolation of amplified and overexpressed DNA sequences from Adriamycin-resistant human breast cancer cells. / Fairchild, C. R.; Ivy, S. P.; Kao-Shan, C. S.; Whang-Peng, J.; Rosen, N.; Israel, M. A.; Melera, P. W.; Cowan, K. H.; Goldsmith, M. E.

於: Cancer Research, 卷 47, 編號 19, 01.01.1987, p. 5141-5148.

研究成果: 雜誌貢獻文章

Fairchild, CR, Ivy, SP, Kao-Shan, CS, Whang-Peng, J, Rosen, N, Israel, MA, Melera, PW, Cowan, KH & Goldsmith, ME 1987, 'Isolation of amplified and overexpressed DNA sequences from Adriamycin-resistant human breast cancer cells', Cancer Research, 卷 47, 編號 19, 頁 5141-5148.
Fairchild CR, Ivy SP, Kao-Shan CS, Whang-Peng J, Rosen N, Israel MA 等. Isolation of amplified and overexpressed DNA sequences from Adriamycin-resistant human breast cancer cells. Cancer Research. 1987 1月 1;47(19):5141-5148.
Fairchild, C. R. ; Ivy, S. P. ; Kao-Shan, C. S. ; Whang-Peng, J. ; Rosen, N. ; Israel, M. A. ; Melera, P. W. ; Cowan, K. H. ; Goldsmith, M. E. / Isolation of amplified and overexpressed DNA sequences from Adriamycin-resistant human breast cancer cells. 於: Cancer Research. 1987 ; 卷 47, 編號 19. 頁 5141-5148.
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title = "Isolation of amplified and overexpressed DNA sequences from Adriamycin-resistant human breast cancer cells",
abstract = "An MCF-7 human breast cancer cell line was selected which was 200-fold more resistant to Adriamycin than the wild type cell line. This Adriamycin-resistant (Adr(R)) cell line exhibited a multidrug-resistant phenotype and was cross-resistant to a wide range of antineoplastic agents including Vinca alkaloids, anthracyclines, and epipodophyllotoxins. Cytogenetic analysis of the Adr(R) cell line showed the presence of homogeneously staining regions on several chromosomes which were not present in the parental cell line. Using the technique of in-gel renaturation, DNA sequences which were amplified 50- to 100-fold in the Adr(R) cell line and which covered a total of over 140 kilobases were isolated. In addition, Adr(R) cells were found to contain amplified and overexpressed sequences which were homologous to hamster P-glycoprotein gene sequences. A hamster cDNA P-glycoprotein gene probe was used to screen a λgt10 cDNA library made from human Adr(R) cell line mRNA and human cDNA sequences homologous to the P-glycoprotein gene were isolated. Hybridization studies with the cloned human cDNA (pADR1) showed that the Adr(R) MCF-7 cell line contained a 60-fold amplification of this DNA sequence and that polyadenylated mRNA from the Adr(R) cell line contained a 4.8-kilobase transcript which was overexpressed 45-fold. There was a direct correlation between DNA and RNA copy number of this sequence and level of resistance among several MCF-7 Adriamycin-resistant cell lines. In situ hybridization studies demonstrated that the human P-glycoprotein gene sequence was found on chromosome 7q21.1 in normal human lymphocytes and that amplified DNA sequences isolated from the Adr(R) MCF-7 cells by the in-gel hybridization technique were linked to the human P-glycoprotein sequences in the homogeneously staining regions in the Adr(R) cells.",
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AU - Kao-Shan, C. S.

AU - Whang-Peng, J.

AU - Rosen, N.

AU - Israel, M. A.

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AU - Cowan, K. H.

AU - Goldsmith, M. E.

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N2 - An MCF-7 human breast cancer cell line was selected which was 200-fold more resistant to Adriamycin than the wild type cell line. This Adriamycin-resistant (Adr(R)) cell line exhibited a multidrug-resistant phenotype and was cross-resistant to a wide range of antineoplastic agents including Vinca alkaloids, anthracyclines, and epipodophyllotoxins. Cytogenetic analysis of the Adr(R) cell line showed the presence of homogeneously staining regions on several chromosomes which were not present in the parental cell line. Using the technique of in-gel renaturation, DNA sequences which were amplified 50- to 100-fold in the Adr(R) cell line and which covered a total of over 140 kilobases were isolated. In addition, Adr(R) cells were found to contain amplified and overexpressed sequences which were homologous to hamster P-glycoprotein gene sequences. A hamster cDNA P-glycoprotein gene probe was used to screen a λgt10 cDNA library made from human Adr(R) cell line mRNA and human cDNA sequences homologous to the P-glycoprotein gene were isolated. Hybridization studies with the cloned human cDNA (pADR1) showed that the Adr(R) MCF-7 cell line contained a 60-fold amplification of this DNA sequence and that polyadenylated mRNA from the Adr(R) cell line contained a 4.8-kilobase transcript which was overexpressed 45-fold. There was a direct correlation between DNA and RNA copy number of this sequence and level of resistance among several MCF-7 Adriamycin-resistant cell lines. In situ hybridization studies demonstrated that the human P-glycoprotein gene sequence was found on chromosome 7q21.1 in normal human lymphocytes and that amplified DNA sequences isolated from the Adr(R) MCF-7 cells by the in-gel hybridization technique were linked to the human P-glycoprotein sequences in the homogeneously staining regions in the Adr(R) cells.

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